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Raimund Dietze Anna Starzinski-Powitz Georgios Scheiner-Bobis Hans-Rudolf Tinneberg Ivo Meinhold-Heerlein Lutz Konrad 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2018,1863(11):1369-1377
Extracellular lysophosphatidic acid (LPA) and the G-protein-coupled LPA receptors (LPAR) are involved in cell migration and invasion and found in the human endometrium. However, underlying mechanisms resulting in cellular invasion have been rarely investigated. We used stromal endometrial T-HESC, epithelial endometriotic 12Z, 49Z and Ishikawa cells. Interestingly, proliferation of T-HESC cells was strongly increased after LPA treatment, whereas the epithelial cell lines only showed a moderate increase. LPA increased invasion of 12Z and 49Z strongly and significantly. The LPAR inhibitor Ki16425 (LPAR1/3) attenuated significantly LPA-induced invasiveness of 12Z, which was confirmed by LPAR1 and LPAR3 siRNAs, showing that both LPA receptors contribute to invasiveness of 12Z cells. Investigation of cell invasion with an antibody-based protease array revealed mainly differences in cathepsins and especially cathepsin B between 12Z compared to the less invasive Ishikawa. Stimulation with LPA showed a time- and dose-dependent increased secretion of cathepsin B which was inhibited by the Gq inhibitor YM-254890 and Gi/o inhibitor pertussis toxin in the 12Z cells, again highlighting the importance of LPAR1/3. The activity of intracellular and secreted cathepsin B was significantly upregulated in LPA-treated samples. Inhibition of cathepsin B with the specific inhibitor CA074 significantly reduced LPA-increased invasion of 12Z. Our results reveal a novel role of LPA-mediated secretion of cathepsin B which stimulated invasion of endometriotic epithelial cells mainly via LPAR1 and LPAR3. These findings may deepen our understanding how endometriotic cells invade into ectopic sites, and provide new insights into the role of LPA and cathepsin B in cellular invasion. 相似文献
23.
Enantioselective Transformation of α-Hexachlorocyclohexane by the Dehydrochlorinases LinA1 and LinA2 from the Soil Bacterium Sphingomonas paucimobilis B90A 下载免费PDF全文
Mrutyunjay Suar Andrea Hauser Thomas Poiger Hans-Rudolf Buser Markus D. Müller Charu Dogra Vishakha Raina Christof Holliger Jan Roelof van der Meer Rup Lal Hans-Peter E. Kohler 《Applied microbiology》2005,71(12):8514-8518
Sphingomonas paucimobilis B90A contains two variants, LinA1 and LinA2, of a dehydrochlorinase that catalyzes the first and second steps in the metabolism of hexachlorocyclohexanes (R. Kumari, S. Subudhi, M. Suar, G. Dhingra, V. Raina, C. Dogra, S. Lal, J. R. van der Meer, C. Holliger, and R. Lal, Appl. Environ. Microbiol. 68:6021-6028, 2002). On the amino acid level, LinA1 and LinA2 were 88% identical to each other, and LinA2 was 100% identical to LinA of S. paucimobilis UT26. Incubation of chiral α-hexachlorocyclohexane (α-HCH) with Escherichia coli BL21 expressing functional LinA1 and LinA2 S-glutathione transferase fusion proteins showed that LinA1 preferentially converted the (+) enantiomer, whereas LinA2 preferred the (−) enantiomer. Concurrent formation and subsequent dissipation of β-pentachlorocyclohexene enantiomers was also observed in these experiments, indicating that there was enantioselective formation and/or dissipation of these enantiomers. LinA1 preferentially formed (3S,4S,5R,6R)-1,3,4,5,6-pentachlorocyclohexene, and LinA2 preferentially formed (3R,4R,5S,6S)-1,3,4,5,6-pentachlorocyclohexene. Because enantioselectivity was not observed in incubations with whole cells of S. paucimobilis B90A, we concluded that LinA1 and LinA2 are equally active in this organism. The enantioselective transformation of chiral α-HCH by LinA1 and LinA2 provides the first evidence of the molecular basis for the changed enantiomer composition of α-HCH in many natural environments. Enantioselective degradation may be one of the key processes determining enantiomer composition, especially when strains that contain only one of the linA genes, such as S. paucimobilis UT26, prevail. 相似文献
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Haloalkane Dehalogenase LinB Is Responsible for β- and δ-Hexachlorocyclohexane Transformation in Sphingobium indicum B90A 下载免费PDF全文
Poonam Sharma Vishakha Raina Rekha Kumari Shweta Malhotra Charu Dogra Hansi Kumari Hans-Peter E. Kohler Hans-Rudolf Buser Christof Holliger Rup Lal 《Applied microbiology》2006,72(9):5720-5727
Incubation of resting cells of Sphingobium indicum B90A, Sphingobium japonicum UT26, and Sphingobium francense Sp+ showed that they were able to transform β- and δ-hexachlorocyclohexane (β- and δ-HCH, respectively), the most recalcitrant hexachlorocyclohexane isomers, to pentachlorocyclohexanols, but only resting cells of strain B90A could further transform the pentachlorocyclohexanol intermediates to the corresponding tetrachlorocyclohexanediols. Moreover, experiments with resting cells of Escherichia coli expressing the LinB proteins of strains B90A, UT26, and Sp+ indicated that LinB was responsible for these transformations. Purified LinB proteins from all three strains also effected the formation of the respective pentachlorocyclohexanols. Although the three LinB enzymes differ only marginally with respect to amino acid sequence, they showed interesting differences with respect to substrate specificity. When LinB from strain B90A was incubated with β- and δ-HCH, the pentachlorocyclohexanol products were further transformed and eventually disappeared from the incubation mixtures. In contrast, the LinB proteins from strains UT26 and Sp+ could not catalyze transformation of the pentachlorocyclohexanols, and these products accumulated in the incubation mixture. A mutant of strain Sp+ lacking linA and linB did not degrade any of the HCH isomers, including β-HCH, and complementation of this mutant by linB from strain B90A restored the ability to degrade β- and δ-HCH. 相似文献
27.
Neurons generate spikes reliably with millisecond precision if driven by a fluctuating current—is it then possible to predict the spike timing knowing the input? We determined parameters of an adapting threshold model using data recorded in vitro from 24 layer 5 pyramidal neurons from rat somatosensory cortex, stimulated intracellularly by a fluctuating current simulating synaptic bombardment in vivo. The model generates output spikes whenever the membrane voltage (a filtered version of the input current) reaches a dynamic threshold. We find that for input currents with large fluctuation amplitude, up to 75% of the spike times can be predicted with a precision of ±2 ms. Some of the intrinsic neuronal unreliability can be accounted for by a noisy threshold mechanism. Our results suggest that, under random current injection into the soma, (i) neuronal behavior in the subthreshold regime can be well approximated by a simple linear filter; and (ii) most of the nonlinearities are captured by a simple threshold process. 相似文献
28.
Andreas Fließbach Manuel Winkler Matthias P. Lutz Hans-Rudolf Oberholzer Paul Mäder 《Microbial ecology》2009,57(4):611-623
Pseudomonas fluorescens strains are used in agriculture as plant growth-promoting rhizobacteria (PGPR). Nontarget effects of released organisms should
be analyzed prior to their large-scale use, and methods should be available to sensitively detect possible changes in the
environments the organism is released to. According to ecological theory, microbial communities with a greater diversity should
be less susceptible to disturbance by invading organisms. Based on this principle, we laid out a pot experiment with field-derived
soils different in their microbial biomass and activity due to long-term management on similar parent geological material
(loess). We investigated the survival of P. fluorescens CHA0 that carried a resistance toward rifampicine and the duration of potential changes of the soil microflora caused by
the inoculation with the bacterium at the sowing date of spring wheat. Soil microbial biomass (C
mic, N
mic) basal soil respiration (BR), qCO2, dehydrogenase activity (DHA), bacterial plate counts, mycorrhiza root colonization, and community level substrate utilization
were analyzed after 18 and 60 days. At the initial stage, soils were clearly different with respect to most of the parameters
measured, and a time-dependent effect between the first and the second set point were attributable to wheat growth and the
influence of roots. The effect of the inoculum was small and merely transient, though significant long-term changes were found
in soils with a relatively low level of microbial biomass. Community level substrate utilization as an indicator of changes
in microbial community structure was mainly changed by the growth of wheat, while other experimental factors were negligible.
The sensitivity of the applied methods to distinguish the experimental soils was in decreasing order N
mic, DHA, C
mic, and qCO2. Besides the selective enumeration of P. fluorescens CHA0 rif+, which was only found in amended soils, methods to distinguish the inoculum effect were DHA, C
mic, and the ratio of C
mic to N
mic. The sampling time was most sensitively indicated by N
mic, DHA, C
mic, and qCO2. Our data support the hypothesis—based on ecosystem theory—that a rich microflora is buffering changes due to invading species.
In other words, a soil-derived bacterium was more effective in a relatively poor soil than in soils that are rich in microorganisms. 相似文献
29.
Hans-Rudolf Weiss Stefano Negrini Manuel Rigo Tomasz Kotwicki Martha C Hawes Theodoros B Grivas Toru Maruyama Franz Landauer 《Scoliosis》2006,1(1):1-5
We report a 15-year-old girl who presented with spinal malsegmentation, associated with other skeletal anomalies. The spinal malsegmentation was subsequently discovered to be part of the spondylocarpotarsal synostosis syndrome. In addition, a distinctive craniocervical malformation was identified, which included atlanto-axial rotatory fixation. The clinical and the radiographic findings are described, and we emphasise the importance of computerised tomography to characterize the craniocervical malformation complex. To the best of our knowledge, this is the first clinical report of a child with spondylocarpotarsal synostosis associated with atlanto-axial rotatory fixation. 相似文献
30.